A protein sequencer facility is requested for UCSD. This facility will consist of a high sensitivity gs phase sequencer (prototype developed at CalTech) and two HPLC machines (commercially available) to analyze the amino acid derivatives generated by the sequencer. The uses of this facility for this campus as proposed by the investigators, fall into several broad categories. A. The sequencing of the N-terminal portion of small amounts of highly purified proteins is proposed as the necessary first step in the construction of oligonucleotide probes and eventual cloning of the genes for these proteins. These proteins are the human HGPRT, human B-galactosidase, a-fucosidase and hexosaminidase A&B; human G-6-dehydrogenase (Kaplan); bacterial virus protein TF1, and Clostridia hydrogenase, and ornithine decarboyxlase and succinate dehydrogenase from mammalian cell cultures. B. The determination of mutationally induced amino acid substitutions in the primary structure of two proteins will be undertaken, yeast alcohol dehydrogenase, and subunit #9 of the mitochondrial ATPase of Neurospora. C. The primary structure will be determined of integral membrane proteins, such as mannitol Enz II of the phosphotransferase system of E. coli (Saier) and the F0 and F1 portions of the ATPase complexes of E. coli and beef heart mitochondria. D. The primary structure will be determined for the enzymes in the tryptophan pathway, and for a key protein in bones, BGP, and for cAMP dependent protein kinase.